Potential health impacts of increasing the cigarette tax in Viet Nam.
Even after the implementation of MPOWER, Viet Nam's cigarette tax remains only 36% of the retail price. This falls short of the World Health Organization's (WHO's) recommendation that the tax share be 70% of the retail price. To assess the health impact of different levels of cigarette taxation by estimating the impact on reducing tobacco consumption and smoking-attributable deaths (SADs). A static model was developed based on a previous study by Jha et al. The model calculates the reduction in tobacco consumption, estimating the number of SADs averted and savings in health-related costs. The scenarios for increasing cigarette taxes were aligned with the Viet Nam Tobacco Tax Simulation Model developed by the WHO and the Vietnamese Ministry of Finance. Four scenarios proposed by the Ministry of Finance and the Ministry of Health would prevent an estimated 63 339-581 165 SADs, equivalent to mortality costs of US$577-5296 million. In the ideal scenario, 6 258 361 SADs would be prevented and would save US$57.0 billion. Future young smokers would see the greatest health benefits of increasing the cigarette tax. Increasing the cigarette tax could reduce the substantial health impact of tobacco use, and further result in significant financial savings across society. Viet Nam should support efforts to further increase the cigarette tax following WHO recommendations.


Fish oil and inflammatory status alter the n-3 to n-6 balance of the endocannabinoid and oxylipin metabolomes in mouse plasma and tissues.
It is well established that dietary intake of n-3 fatty acids is associated with anti-inflammatory effects, and this has been linked to modulation of the oxylipin and endocannabinoid metabolomes. However, the amount of data on specific tissue effects is limited, and it is not known how inflammation affects this relation. In the present study we systematically explored the combined effects of n-3 fatty acid diets and inflammation on the in vivo endocannabinoid and oxylipin metabolomes using a multicompartment, detailed targeted lipidomics approach. Male C57BL/6 mice received diets containing 0, 1, or 3 % w/w fish oil (FO) for 6 weeks, after which 2 mg/kg LPS or saline was administered i.p. Levels of endocannabinoids/N-acylethanolamines (NAEs) and oxylipins, covering n-3 and n-6 fatty acid derived compounds, were determined in plasma, liver, ileum and adipose tissue using LC-MS/MS. FO generally increased 'n-3' NAEs and oxylipins at the expense of compounds derived from other fatty acids, affecting all branches of the oxylipin metabolome. LPS generally increased levels of endocannabinoids/NAEs and oxylipins, with opposing effects across plasma and tissues. Multivariate data analysis revealed that separation between diet groups in the saline treated groups was primarily explained by decreases in other than n-3 derived compounds. In the LPS treated groups, the separation was primarily explained by increases in n-3 derived compounds. In conclusion, FO caused marked changes in the n-3 to n-6 balance of the endocannabinoid and oxylipin metabolomes, with specific effects depending on inflammatory status. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11306-012-0421-9) contains supplementary material, which is available to authorized users.


Complex regional pain syndrome 1--the Swiss cohort study.
Little is known about the course of Complex Regional Pain Syndrome 1 and potential factors influencing the course of this disorder over time. The goal of this study is a) to set up a database with patients suffering from suspected CRPS 1 in an initial stadium, b) to perform investigations on epidemiology, diagnosis, prognosis, and socioeconomics within the database and c) to develop a prognostic risk assessment tool for patients with CRPS 1 taking into account symptomatology and specific therapies. Prospective cohort study. Patients suffering from a painful swelling of the hand or foot which appeared within 8 weeks after a trauma or a surgery and which cannot be explained by conditions that would otherwise account for the degree of pain and dysfunction will be included. In accordance with the recommendations of International Classification of Functioning, Disability and Health (ICF model), standardised and validated questionnaires will be used. Patients will be monitored over a period of 2 years at 6 scheduled visits (0 and 6 weeks, 3, 6, 12, and 24 months). Each visit involves a physical examination, registration of therapeutic interventions, and completion of the various study questionnaires. Outcomes involve changes in health status, quality of life and costs/utility. This paper describes the rationale and design of patients with CRPS 1. Ideally, potential risk factors may be identified at an early stage in order to initiate an early and adequate treatment in patients with increased risk for delayed recovery.


Using blogging tools to help individuals record their experiences: an exploration and review of two commercial web applications in the Netherlands.
Blogs, short for "web logs," together with podcasts and wikis are currently important foci of general internet research. These three applications are part of the larger body of next-generation communication applications that comprises "Web 2.0." Within the specific area of health care, however, little attention has been devoted to understanding these technologies and how they are being used by lay health publics. In this article, I will discuss the emergent findings from a new project that looks at blogging interfaces as potential tools for disease prevention and health promotion. I use a literature review combined with "front stage" web analyses of two cases and interviews with the supporting institutions for these sites to discuss the relevant informatics questions that arise with respect to these applications. I further introduce the idea of "goal-oriented" blogging that is found in the first case study. Because this research project is still in preliminary phases, this should be viewed as an exploration into the topic and work in progress. In addition to raising questions, I will outline the important subsequent research steps.


BMI-1 promotes ewing sarcoma tumorigenicity independent of CDKN2A repression.
Deregulation of the polycomb group gene BMI-1 is implicated in the pathogenesis of many human cancers. In this study, we have investigated if the Ewing sarcoma family of tumors (ESFT) expresses BMI-1 and whether it functions as an oncogene in this highly aggressive group of bone and soft tissue tumors. Our data show that BMI-1 is highly expressed by ESFT cells and that, although it does not significantly affect proliferation or survival, BMI-1 actively promotes anchorage-independent growth in vitro and tumorigenicity in vivo. Moreover, we find that BMI-1 promotes the tumorigenicity of both p16 wild-type and p16-null cell lines, demonstrating that the mechanism of BMI-1 oncogenic function in ESFT is, at least in part, independent of CDKN2A repression. Expression profiling studies of ESFT cells following BMI-1 knockdown reveal that BMI-1 regulates the expression of hundreds of downstream target genes including, in particular, genes involved in both differentiation and development as well as cell-cell and cell-matrix adhesion. Gain and loss of function assays confirm that BMI-1 represses the expression of the adhesion-associated basement membrane protein nidogen 1. In addition, although BMI-1 promotes ESFT adhesion, nidogen 1 inhibits cellular adhesion in vitro. Together, these data support a pivotal role for BMI-1 ESFT pathogenesis and suggest that its oncogenic function in these tumors is in part mediated through modulation of adhesion pathways.


Founding fathers of light scattering and surface-enhanced Raman scattering.
One can view our comprehension of surface-enhanced Raman scattering, particularly that by colloidal dispersions of metal sols, as the merging of two traditions in light-scattering theory and practice. One of these originated with Michael Faraday's work on brilliantly colored metal sols, which was taken up by Richard Zsigmondy and then by Gustav Mie, who accounted for the colors by electromagnetic-scattering theory. The other tradition starts with John Tyndall's work with aerosols, which stimulated Lord Rayleigh's entry into the field. Lord Rayleigh was perplexed by observations made with sulfur hydrosols, which in turn were explored by C. V. Raman. Raman's extensive work in light scattering led to his subsequent discovery of the Raman effect. These two traditions were then intertwined when it was shown that the same physical effect that caused Faraday's sols to exhibit their brilliant colors was also the origin of the enhancement of Raman signals from molecules adsorbed on the metal particles that compose these sols.


A CpG-loaded tumor cell vaccine induces antitumor CD4+ T cells that are effective in adoptive therapy for large and established tumors.
We designed a whole tumor cell vaccine by "loading" lymphoma tumor cells with CG-enriched oligodeoxynucleotide (CpG), a ligand for the Toll-like receptor 9 (TLR9). CpG-loaded tumor cells were phagocytosed, delivering both tumor antigen(s) and the immunostimulatory CpG molecule to antigen-presenting cells (APCs). These APCs then expressed increased levels of costimulatory molecules and induced T-cell immunity. TLR9 was required in the APCs but not in the CpG-loaded tumor cell. We demonstrate that T cells induced by this vaccine are effective in adoptive cellular therapy for lymphoma. T cells from vaccinated mice transferred into irradiated, syngeneic recipients protected against subsequent lymphoma challenge and, remarkably, led to regression of large and established tumors. This therapeutic effect could be transferred by CD4(+) but not by CD8(+) T cells. A CpG-loaded whole-cell vaccination is practical and has strong potential for translation to the clinical setting. It is currently being tested in a clinical trial of adoptive immunotherapy for mantle-cell lymphoma.


Assessment of changes in general health status using the short-form 36 questionnaire 1 year following coronary artery bypass grafting.
The problem addressed in the study was to gain a greater understanding of the health benefits of coronary artery bypass grafting (CABG). The purpose of the study was to assess general health status, using the short-form (SF)-36 questionnaire, approximately 12 months following CABG, and to document any associations between pre-operative health status, level of social support, coronary artery disease (CAD) risk factors, CAD symptom severity and post-operative health status. The study was prospective and observational in design and included assessments at two time points, namely pre-operatively in a hospital outpatient department (1995-1996) and post-operatively at home (1996-1997). Two hundred and fourteen patients awaiting elective CABG were recruited a month before the expected date of operation. Pre-operative assessment included: (1), severity of symptoms; (2), CAD risk factors; (3), SF-36 questionnaire; and (4), social activities questionnaire. Post-operative assessment measured health status using the SF-36 instrument (mean, 16.4 months). Correlation and multiple linear regression analyses were used to identify factors associated with improved health status following CABG. Two hundred and fourteen patients were assessed pre-operatively and underwent CABG. There was a 4.8% 30-day mortality rate, and 183 patients were followed for a mean of 16.4 months after CABG. SF-36 scores following CABG were improved across all of the eight domains (P<0.001). A higher social network score and higher pre-operative health status were associated with improved health status. Patients with lower health levels (SF-36 scores) prior to CABG were less likely to gain improvement in health (SF-36 scores) following CABG. Lower SF-36 scores following operation were influenced by the presence of diabetes mellitus, cigarette smoking, younger age, a high socio-economic deprivation category and higher alcohol intake. Many patients had uncorrected CAD risk factors at pre-operative assessment. The SF-36 instrument was shown to be a useful and sensitive tool to assess differences and changes in the general health status of patients before and following CABG. High levels of social support were associated with improved health status post-operatively. Lower pre-operative general health status, the presence of diabetes mellitus and cigarette smoking were associated with poorer post-operative general health status.


[Sinonasal fungal infections are not exclusively due to mucorales and Aspergillus!].
Rhino-sinusal infections are serious diseases and possibly lethal. When they are invasive, we easily discuss apergilloses and mucormycoses. The confirmation of the diagnosis of mucormycosis need an extensive surgery for precise histopathological and mycological evaluation. The pathologist may be faced to other rare mycoses such as phaeohyphomycoses, which present different morphological features than mucormycoses and Aspergillus. Once the diagnosis is established, an appropriate antifungal treatment is quickly started. The aim of our work is to report two observations of phaeohyphomycoses, to describe their histopathological features, to discuss complementary diagnostic methods and to present the main differential diagnoses.


Comparison of efficacy and safety of choline fenofibrate (fenofibric acid) to micronized fenofibrate in patients of mixed dyslipidemia: A randomized, open-label, multicenter clinical trial in Indian population.
Choline fenofibrate is a newly developed choline salt of fenofibric acid, which is more hydrophilic than fenofibrate. This study was initiated to evaluate the safety and efficacy of choline fenofibrate in comparison to micronized fenofibrate among Indian patients of mixed dyslipidemia. A multicenter, open-label, randomized, active controlled, comparative, parallel group study was conducted at around 10 centers spread all across the country. Mixed dyslipidemia patients (serum triglycerides [TG] levels between 150 and 500 mg/dl), aged 18-70 years and taking stable statin dose for 8 weeks were randomized to choline fenofibrate 135 mg delayed release tablets and micronized fenofibrate 160 mg tablets once daily for 12 weeks. The primary endpoint of the study was percentage change in serum TG level at the end of 12 weeks. A total of 226 patients were enrolled in this study, of which 116 patients were administered choline fenofibrate and 110 patients were administered micronized fenofibrate. At the end of 12 weeks, there was a significant reduction in TG level (34.24% in choline fenofibrate group and 38.13% reduction in micronized fenofibrate group). However, the difference between group was not statistically different (P = 0.471). Similarly, there was a significant increase in high-density lipoprotein cholesterol at the end of 12 weeks (10% increase in choline fenofibrate group and 9% increase in micronized fenofibrate group); but the difference between the group was not statistically significant (P = 0.598). Both the treatment was safe and well tolerated. Choline fenofibrate delayed release 135 mg is as safe and effective as 160 mg of micronized fenofibrate in Indian patients with mixed dyslipidemia.


Tubulin Polymerization Promoting Protein, Ringmaker, and MAP1B Homolog Futsch Coordinate Microtubule Organization and Synaptic Growth.
Drosophila Ringmaker (Ringer) is homologous to the human Tubulin Polymerization Promoting Proteins (TPPPs) that are implicated in the stabilization and bundling of microtubules (MTs) that are particularly important for neurons and are also implicated in synaptic organization and plasticity. No in vivo functional data exist that have addressed the role of TPPP in synapse organization in any system. Here, we present the phenotypic and functional characterization of ringer mutants during Drosophila larval neuromuscular junction (NMJ) synaptic development. ringer mutants show reduced synaptic growth and transmission and display phenotypic similarities and genetic interactions with the Drosophila homolog of vertebrate Microtubule Associated Protein (MAP)1B, futsch. Immunohistochemical and biochemical analyses show that individual and combined loss of Ringer and Futsch cause a significant reduction in MT loops at the NMJs and reduced acetylated-tubulin levels. Presynaptic over-expression of Ringer and Futsch causes elevated levels of acetylated-tubulin and significant increase in NMJ MT loops. These results indicate that Ringer and Futsch regulate synaptic MT organization in addition to synaptic growth. Together our findings may inform studies on the close mammalian homolog, TPPP, and provide insights into the role of MTs and associated proteins in synapse growth and organization.


Antidepressants for the treatment of abdominal pain-related functional gastrointestinal disorders in children and adolescents.
Abdominal pain-related functional gastrointestinal disorders (FGIDs) are among the most common medical problems in paediatric medicine. Frequently, physicians prescribe antidepressants as a second-line treatment for children and adolescents with FGIDs. To date, the evidence on the benefits and harms of antidepressants for the treatment of abdominal pain-related FGIDs has not been assessed systematically. The primary objectives were to conduct a systematic review to evaluate the efficacy and safety of antidepressants for the treatment of abdominal pain-related FGIDs in children and adolescents. We searched The Cochrane Library, PubMed, EMBASE, IPA, CINAHL, PsycINFO, ISI Web of Science, Biosis Previews and the International Clinical Trials Registry Platform of the World Health Organization with appropriate filters (from inception to January 31, 2011). For efficacy we included double-blind, randomised controlled trials (RCTs) of antidepressants for treatment of abdominal pain-related FGIDs in children and adolescents 18 years or younger. Open-label and uncontrolled experimental studies, as well as observational studies were eligible for the assessment of harms. The minimum study duration was 4 weeks. The minimum study size was 30 participants. Two authors independently assessed all abstracts and full text articles, and rated the risk of bias for included studies. Data were extracted independently by one author and checked for accuracy by another author. Data were analysed using RevMan 5. Two RCTs (123 participants), both using amitriptyline, met the pre-specified inclusion criteria. These studies provided mixed findings on the efficacy of amitriptyline for the treatment of abdominal pain-related FGIDs. The larger, publicly-funded study reported no statistically significant difference in efficacy between amitriptyline and placebo in 90 children and adolescents with FGIDs after 4 weeks of treatment. On intention-to-treat (ITT)- analysis, 59% of the children reported feeling better in the amitriptyline group compared with 53% in the placebo group (RR 1.12; 95% CI: 0.77 to 1.63; P = 0.54). The risk of bias for this study was rated as low.The second RCT enrolled 33 adolescents with irritable bowel syndrome. Patients receiving amitriptyline experienced greater improvements in the primary outcome, overall quality of life, at weeks 6, 10, and 13 compared with those on placebo (P= 0.019, 0.004, and 0.013, respectively). No effect estimates were calculated for the quality of life outcome because mean quality of life scores and standard deviations were not reported. For most secondary outcomes no statistically significant differences between amitriptyline and placebo could be detected. The risk of bias for this study was rated as unclear for most items. However, it was rated as high for other bias due to multiple testing. The results of this study should be interpreted with caution due to the small number of patients and multiple testing.The larger study reported mild adverse events including fatigue, rash and headache and dizziness. On ITT analysis, 4% of the amitriptyline group experienced at least one adverse event compared to 2% of the placebo group. There was no statistically significant difference in the proportion of patients who experienced at least one adverse event (RR 1.91; 95% CI 0.18 to 20.35; P = 0.59). The smaller study reported no adverse events. The methods of adverse effects assessment was poorly reported in both studies and no clear conclusions on the risks of harms of amitriptyline can be drawn. Clinicians must be aware that for the majority of antidepressant medications no evidence exists that supports their use for the treatment of abdominal pain-related FGIDs in children and adolescents. The existing randomised controlled evidence is limited to studies on amitriptyline and revealed no statistically significant differences between amitriptyline and placebo for most efficacy outcomes. Amitriptyline does not appear to provide any benefit for the treatment of FGIDs in children and adolescents. Studies in children with depressive disorders have shown that antidepressants can lead to substantial, sometimes life-threatening adverse effects. Until better evidence evolves, clinicians should weigh the potential benefits of antidepressant treatment against known risks of antidepressants in paediatric patients.


Comparing adult hippocampal neurogenesis in mammalian species and orders: influence of chronological age and life history stage.
Adult hippocampal neurogenesis is a prominent event in rodents. In species with longer life expectancies, newly born cells in the adult dentate gyrus of the hippocampal formation are less abundant or can be completely absent. Several lines of evidence indicate that the regulatory mechanisms of adult neurogenesis differ between short- and long-lived mammals. After a critical appraisal of the factors and problems associated with comparing different species, we provide a quantitative comparison derived from seven laboratory strains of mice (BALB, C57BL/6, CD1, outbred) and rats (F344, Sprague-Dawley, Wistar), six other rodent species of which four are wild-derived (wood mouse, vole, spiny mouse and guinea pig), three non-human primate species (marmoset and two macaque species) and one carnivore (red fox). Normalizing the number of proliferating cells to total granule cell number, we observe an overall exponential decline in proliferation that is chronologically equal between species and orders and independent of early developmental processes and life span. Long- and short-lived mammals differ with regard to major life history stages; at the time points of weaning, age at first reproduction and average life expectancy, long-lived primates and foxes have significantly fewer proliferating cells than rodents. Although the database for neuronal differentiation is limited, we find indications that the extent of neuronal differentiation is subject to species-specific selective adaptations. We conclude that absolute age is the critical factor regulating cell genesis in the adult hippocampus of mammals. Ontogenetic and ecological factors primarily influence the regulation of neuronal differentiation rather than the rate of cell proliferation.


Regulation of the Ets-related transcription factor Elf-1 by binding to the retinoblastoma protein.
The retinoblastoma gene product (Rb) is a nuclear phosphoprotein that regulates cell cycle progression. Elf-1 is a lymphoid-specific Ets transcription factor that regulates inducible gene expression during T cell activation. In this report, it is demonstrated that Elf-1 contains a sequence motif that is highly related to the Rb binding sites of several viral oncoproteins and binds to the pocket region of Rb both in vitro and in vivo. Elf-1 binds exclusively to the underphosphorylated form of Rb and fails to bind to Rb mutants derived from patients with retinoblastoma. Co-immunoprecipitation experiments demonstrated an association between Elf-1 and Rb in resting normal human T cells. After T cell activation, the phosphorylation of Rb results in the release of Elf-1, which is correlated temporally with the activation of Elf-1-mediated transcription. Overexpression of a phosphorylation-defective form of Rb inhibited Elf-1-dependent transcription during T cell activation. These results demonstrate that Rb interacts specifically with a lineage-restricted Ets transcription factor. This regulated interaction may be important for the coordination of lineage-specific effector functions such as lymphokine production with cell cycle progression in activated T cells.


Effect of habitat fragmentation on levels and patterns of genetic diversity in natural populations of the peat moss Polytrichum commune.
Peat bogs represent unique ecosystems that are under particular threat from fragmentation due to peat harvesting, with only 38% of the original peatland in Europe remaining intact and unaffected by peat cutting, drainage and silviculture. In this study, we have used microsatellite markers to determine levels and patterns of genetic diversity in both cut and uncut natural populations of the peat moss Polytrichum commune. Overall diversity levels suggest that there is more genetic variation present than had previously been assumed for bryophytes. Despite this, diversity values from completely cut bogs were found to be lower than those from uncut peatlands (average 0.729 versus 0.880). In addition, the genetic diversity was more highly structured in the cut populations, further suggesting that genetic drift is already affecting genetic diversity in peat bogs subjected to fragmentation.


The JAK1/JAK2- inhibitor ruxolitinib inhibits mast cell degranulation and cytokine release.
Mastocytosis is characterized by the accumulation of aberrant mast cells (MC). Patients suffering from mastocytosis suffer from a wide range of symptoms due to increased levels of MC mediators. It would therefore be of great benefit to inhibit MC mediator release. However, to date there are few drugs available that are known to effectively lower MC mediator levels. The evidence for the involvement of the janus kinase 2 (JAK2)-signal transducer and activation of transcription 5 (STAT5) signalling pathway in MC activation is slowly accumulating. Interference with the JAK2-STAT5 pathway might inhibit MC mediator release. Ruxolitinib, a JAK1/JAK2 inhibitor, indeed decreases symptoms like pruritus and fatigue in patients with myeloproliferative neoplasms. Yet, detailed studies on how ruxolitinib affects human mast cell activity are lacking. To investigate the effect of JAK1/2-inhibition with ruxolitinib in the human mast cell lines LAD2 and HMC1. LAD2 and HMC1 were stimulated with substance P, codeine or the calcium ionophore A23817. The effect of ruxolitinib on mast cell degranulation (via measurement of β-hexosaminidase, histamine release and CD63 membrane expression) and IL-6, IL-13, MCP-1 and TNF-α production was investigated. The involvement of STAT5 activation was explored using the selective STAT5 inhibitor pimozide. Ruxolitinib effectively inhibited codeine- and substance P-induced degranulation in a concentration-dependent manner. Ruxolitinib also significantly inhibited the production of IL-6, TNF-α and MCP-1 as induced by A23817 and substance P. Selective STAT5 inhibition with pimozide resulted in diminished degranulation and inhibition of cytokine production as induced by A23817 and substance P. This study demonstrates that the JAK1/JAK2 inhibitor ruxolitinib can inhibit MCactivity, possibly through prevention of STAT5 activation. This renders the JAK-STAT pathway as an interesting target for therapy to release symptom burden in mastocytosis and many other MC mediator-related diseases.


First Report of Powdery Mildew Caused by Erysiphe viburni on Viburnum tinus in California.
In June 2003, landscape and potted nursery plants of laurustinus (Viburnum tinus) in Monterey County, California, were found to be infected with a powdery mildew. White, ectophytic mycelial and conidial growth were present primarily on adaxial leaf sides with only sparse growth on abaxial surfaces. Severely infected leaves were buckled and slightly twisted. Affected leaf tissue exhibited slight purple-to-brown discoloration. Appressoria were opposite and lobed. Conidia were produced singly, cylindrical in shape, and measured 31 to 42 × 14 to 19 μm. No fibrosin bodies were observed in the conidia, and the conidia germinated at the ends. Ascomata were not observed. The fungus was identified as Erysiphe (section Microsphaera) viburni Duby (= Microsphaera sparsa Howe = Microsphaera penicillata [M. sparsa is not completely synonymous with M. penicillata because M. sparsa is defined as only those mildews that attack viburnum, whereas M. penicillata was defined as attacking dogwood, alder, etc.]) (1,2). Pathogenicity was demonstrated by gently pressing infected leaves with abundant sporulation onto recently expanded leaves of four, large, potted laurustinus in standard 5-gallon nursery containers (19 liters). Twelve leaves per plant were inoculated. The plants were incubated in a greenhouse (23 to 25°C). After 9 to 10 days, powdery mildew colonies developed on approximately 50% of the inoculated leaves. Such colonies were morphologically similar to the original fungus. Uninoculated control plants did not develop powdery mildew. Using the same technique, inoculating fully mature leaves of the same plants did not result in disease. To our knowledge, this is the first report of Erysiphe viburni infecting V. tinus in California. At some landscaped areas the powdery mildew was extremely severe, causing plants to take on a whitish appearance and resulting in all new foliage being misshapen. E. viburni has also been reported to infect V. japonicum in California. References: (1) U. Braun. The Powdery Mildews (Erysiphales) of Europe. Gustave Fischer. New York, 1995. (2) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000.


Endothelial overexpression of Fas ligand decreases atherosclerosis in apolipoprotein E-deficient mice.
Fas ligand (FasL) can induce apoptosis in cells bearing the Fas receptor. The role of FasL in the vasculature with regard to atherosclerosis is controversial. This study examined the function of endothelial FasL during atherosclerosis. Transgenic (Tg) mice that specifically overexpress different levels of FasL on vascular endothelial cells were crossed into the apolipoprotein E-knockout background (ApoE-KO) to generate ApoE-KO/FasL-Tg mice. Although plasma cholesterol and triglyceride levels were not different between ApoE-KO/FasL-Tg mice and ApoE-KO mice after 12 weeks of a high-fat diet, overexpression of the FasL transgene significantly reduced atherosclerotic lesion area in aortae by 49%. The reduction of atherosclerotic lesion area was more pronounced in thoracic and abdominal aortae than in the aortic arch, and a 34% reduction in lesion area was observed in aortic root sections from the ApoE-KO/FasL-Tg group compared with the ApoE-KO group. Immunostaining revealed significant decreases in both macrophage and CD8 T-cell accumulation in lesions of ApoE-KO/FasL-Tg mice. ApoE-KO/FasL-Tg mice that express lower levels of endothelial FasL also displayed reduced lesion size, but this reduction was statistically significant at the aortic arch only. Overexpression of endothelial FasL is antiinflammatory and inhibits atherosclerosis under hypercholesterolemic conditions.


Effect of Targeted Pulsed Electromagnetic Field Therapy on Canine Postoperative Hemilaminectomy: A Double-Blind, Randomized, Placebo-Controlled Clinical Trial.
Intervertebral disc disease is one of the leading causes of paralysis in dogs. Pulsed electromagnetic field (PEMF) therapy has been advocated for improving wound healing and pain reduction; however, robust clinical trials are lacking. The present prospective, double-blind, placebo-controlled trial evaluated targeted PEMF therapy administered to 53 client-owned dogs who underwent hemilaminectomy for naturally occurring disk extrusion intervertebral disc disease. The dogs were randomized to receive either targeted PEMF (n = 27) or placebo treatment (n = 28). Wound healing, evaluated by visual analog score and wound evaluation scale, was significantly improved at 6 wk postoperatively in the treatment compared with the control group (P = .010 and .023, respectively). Pain medications were administered less frequently in dogs receiving PEMF treatment during the 7 day postoperative period compared with the control treatment group (P = .010) with codeine administered 1.8 times more frequently in the control group. No untoward effects were recorded in either treatment group. More frequent evaluation of outcome measures with larger patient numbers, as well as histologic samples, may be useful in future studies. Dogs receiving PEMF therapy following postoperative hemilaminectomy demonstrated improved wound scores at 6 wk and reduced mean number of owner-administered pain medications compared with the control group therapy.


Genetic alteration of Keap1 confers constitutive Nrf2 activation and resistance to chemotherapy in gallbladder cancer.
Biliary tract cancer (BTC) is a highly malignant tumor, and identification of effective therapeutic targets to improve prognosis is urgently required. Oncogenic activation of survival genes is important for cancer cells to overcome oxidative stresses induced by their microenvironments that include chronic inflammation or exposure to anticancer drugs. We attempted to examine whether deregulation of Nrf2, a master transcriptional factor of various cytoprotective genes against oxidative stress, plays a role in the carcinogenesis of BTC. We screened genetic alteration of Keap1, a negative regulator of Nrf2, in BTC including tumors originated from gallbladder and extra- and intrahepatic bile ducts. Functional analysis of cancer-related mutant Keap1 in Nrf2 repression and the association between Nrf2 activation and resistance to 5-fluorouracil (5-FU) were investigated. Recurrent (in 1/11 cell lines and 6/53 primary tumors) Keap1 gene alterations were observed in BTC and were especially frequent (4/13, 30.7%) in gallbladder cancer (GBC). These alterations led to a considerable loss of Nrf2 repression activity, caused constitutive activation of Nrf2, and promoted cell proliferation. Down-regulation of Nrf2 activity by either Keap1 complementation or Nrf2 short interference RNA increased sensitivity to 5-FU in Keap1-altered BTC cells. Keap1 mutation occurs frequently in GBC. Aberrant Nrf2 activation provoked by Keap1 alteration is one of the molecular mechanisms for chemotherapeutic resistance in GBC and will be a novel therapeutic target as an enhancer of sensitivity to 5-FU-based regimens.


Ornithine decarboxylase is a mediator of c-Myc-induced apoptosis.
c-Myc plays a central role in the regulation of cell cycle progression, differentiation, and apoptosis. However, the proteins which mediate c-Myc function(s) remain to be determined. Enforced c-myc expression rapidly induces apoptosis in interleukin-3 (IL-3)-dependent 32D.3 murine myeloid cells following IL-3 withdrawal, and this is associated with the constitutive, growth factor-independent expression of ornithine decarboxylase (ODC), a rate-limiting enzyme of polyamine biosynthesis. Here we have examined the role of ODC in c-Myc-induced apoptosis. Enforced expression of ODC, like c-myc, is sufficient to induce accelerated death following IL-3 withdrawal. ODC induced cell death in a dose-dependent fashion, and alpha-difluoromethylornithine (DFMO), an irreversible inhibitor of ODC enzyme activity, effectively blocked ODC-induced cell death. ODC-induced cell death was due to the induction of apoptosis. We also demonstrate that ODC is a mediator of c-Myc-induced apoptosis. 32D.3-derived c-myc clones have augmented levels of ODC enzyme activity, and their rates of death were also a function of their ODC enzyme levels. Importantly, the rates of death of c-myc clones were inhibited by treatment with DFMO. These findings demonstrate that ODC is an important mediator of c-Myc-induced apoptosis and suggest that ODC mediates other c-Myc functions.


Probing the Morphology and Evolving Dynamics of 3D Printed Nanostructures Using High-Speed Atomic Force Microscopy.
Focused electron beam induced deposition (FEBID) has been demonstrated as a promising solution for synthesizing truly three-dimensional (3D) nanostructures. However, the lack of morphological feedback during growth complicates further development toward higher spatial fabrication precision. Here, we show that by combining in situ high speed atomic force microscopy (HS-AFM) with FEBID, morphologies in multistep fabrication process can be accessed. More importantly, the proposed method enables simultaneous imaging and fabrication operation, which opens new possibilities to investigate evolving mechanical properties of the deposit. The experiments indicate an exponential increase law of the mechanical resistance, meaning that a mechanically stable state establishes around 4 min after deposition.


Effects of intestinal bypass surgery on appetite, food intake, and body weight in obese and lean rats.
Jejunoileal bypass surgery or sham surgery was performed in female rats made obese with ventromedial hypothalamic (VMH) knife cuts, and in lean control rats. After bypass surgery, the VMH rats underate and lost weight until they reached the body weight of the control sham rats, and they then maintained their weight at control levels. Bypass surgery in lean rats produced much smaller reductions in food intake and body weight. Both bypass groups initially consumed less of a sucrose solution and milk diet during 1 h/day tests, but their intakes returned to near normal levels during the second postoperative month. Reconnection of the intestinal tract in the VMH-bypass rats led to renewed hyperphagia and return to obese body weights. A second experiment revealed that bypass surgery reduces food intake and body weight in genetically obese (fatty) rats, but this effect is not as pronounced as that displayed by VMH rats. These results confirm recent clinical observations that reduced appetite and caloric intake are the major causes of the weight loss produced by intestinal bypass surgery.


Concerted or stepwise: how much do free-energy landscapes tell us about the mechanisms of elimination reactions?
The base-catalyzed dehydration of benzene cis-1,2-dihydrodiols is driven by formation of an aromatic product as well as intermediates potentially stabilized by hyperaromaticity. Experiments exhibit surprising shifts in isotope effects, indicating an unusual mechanistic balance on the E2-E1cB continuum. In this study, both 1- and 2-dimensional free energy surfaces are generated for these compounds with various substituents, using density functional theory and a mixed implicit/explicit solvation model. The computational data help unravel hidden intermediates along the reaction coordinate and provide a novel conceptual framework for distinguishing between competing pathways in this and any other system with borderline reaction mechanisms.


Potential of lignocellulosic fractions from Posidonia oceanica to improve barrier and mechanical properties of bio-based packaging materials.
This work reports on the valorization of residues from Posidonia oceanica leaves for the purpose of obtaining lignocellulosic fractions of interest for the development of bio-based materials for food packaging. The lignocellulosic fractions were characterized, thereby confirming the purification of cellulose and showing increases in crystallinity and thermal stability after the consecutive extraction steps. Subsequently, pure lignocellulosic films were obtained and characterized and the pure cellulose film showed the best properties in terms of mechanical performance and water vapor permeability. Finally, composite starch films containing lignocellulosic fractions were developed by melt compounding and characterized. Although the film containing the pure cellulose additive showed the optimum improvement in terms of mechanical properties (with an increase of 85% in the elastic modulus and 38% in the tensile strength), similar water vapor permeability reduction (~40%) was achieved with the least purified fractions, explained by their effect on starch gelatinization, as evidenced by SAXS/WAXS.


Dentin to dentin adhesion using combinations of resin cements and adhesives from different manufacturers - a novel approach.
The aims of this study were to present a novel method to analyse dentin bond strength and to evaluate the bond strength of combining adhesive systems and resin cement from different manufacturers. Human wisdom teeth were ground flat to the dentin on parallel surfaces and axially cut into two parts. Dentin cylinders (Ø 3 mm) were drilled from one half of each tooth. The other half from each tooth was embedded in epoxy resin with the dentin surface exposed. The specimens were ground with silicone carbide paper and the dentin cylinders were cemented onto the dentin surface of the other half of the same tooth.Materials: Resin cement and adhesive systems from three different manufacturers were used in various combinations (n = 8 per group). Cement and adhesive from the same manufacturer served as control. Shear bond strength (SBS) was measured and fracture modes were registered.Results and conclusions: The highest median SBS value was found in a bonding combination between cement and a non-corresponding adhesive (33.1 MPa) and one of the lowest values was found in one of the controls (15.3 MPa). Cohesive fractures were most frequent. The results indicated that combining adhesive and cement from different manufacturers did not compromise the dentin bonding. The novel test method is recommended for evaluating dentin bonding.


The comprehensive assessment of health status in survivors of childhood cancer: application to high-risk acute lymphoblastic leukaemia.
The health status of 69 survivors of high-risk acute lymphoblastic leukaemia (ALL) is assessed using a multi-attribute classification system. Seven attributes are included: sensation, mobility, emotion, cognition, self-care, pain and fertility. Three to five levels of functioning are defined for each attribute. Comprehensive health states are described as a specific combination of seven attribute levels. The system captures combinations of sequelae. The system provides a compact but comprehensive tool for long term follow up of survivors of childhood cancer. The results underscore the cognitive and emotional burdens of morbidity affecting survivors of high-risk ALL.


DNA methylation and hormone receptor status in breast cancer.
We examined whether differences in tumor DNA methylation were associated with more aggressive hormone receptor-negative breast cancer in an ethnically diverse group of patients in the Breast Cancer Care in Chicago (BCCC) study and using data from The Cancer Genome Atlas (TCGA). DNA was extracted from formalin-fixed, paraffin-embedded samples on 75 patients (21 White, 31 African-American, and 23 Hispanic) (training dataset) enrolled in the BCCC. Hormone receptor status was defined as negative if tumors were negative for both estrogen and progesterone (ER/PR) receptors (N = 22/75). DNA methylation was analyzed at 1505 CpG sites within 807 gene promoters using the Illumina GoldenGate assay. Differential DNA methylation as a predictor of hormone receptor status was tested while controlling for false discovery rate and assigned to the gene closest to the respective CpG site. Next, those genes that predicted ER/PR status were validated using TCGA data with respect to DNA methylation (validation dataset), and correlations between CpG methylation and gene expression were examined. In the training dataset, 5.7 % of promoter mean methylation values (46/807) were associated with receptor status at P < 0.05; for 88 % of these (38/46), hypermethylation was associated with receptor-positive disease. Hypermethylation for FZD9, MME, BCAP31, HDAC9, PAX6, SCGB3A1, PDGFRA, IGFBP3, and PTGS2 genes most strongly predicted receptor-positive disease. Twenty-one of 24 predictor genes from the training dataset were confirmed in the validation dataset. The level of DNA methylation at 19 out 22 genes, for which gene expression data were available, was associated with gene activity. Higher levels of promoter methylation strongly correlate with hormone receptor positive status of breast tumors. For most of the genes identified in our training dataset as ER/PR receptor status predictors, DNA methylation correlated with stable gene expression level. The predictors performed well when evaluated on independent set of samples, with different racioethnic distribution, thus providing evidence that this set of DNA methylation biomarkers will likely generalize to prospective patient samples.


Listeria monocytogenes strains show large variations in competitive growth in mixed culture biofilms and suspensions with bacteria from food processing environments.
Interactions and competition between resident bacteria in food processing environments could affect their ability to survive, grow and persist in microhabitats and niches in the food industry. In this study, the competitive ability of L. monocytogenes strains grown together in separate culture mixes with other L. monocytogenes (L. mono mix), L. innocua (Listeria mix), Gram-negative bacteria (Gram- mix) and with a multigenera mix (Listeria + Gram- mix) was investigated in biofilms on stainless steel and in suspensions at 12 °C. The mixed cultures included resident bacteria from processing surfaces in meat and salmon industry represented by L. monocytogenes (n = 6), L. innocua (n = 5) and Gram-negative bacteria (n = 6; Acinetobacter sp., Pseudomonas fragi, Pseudomonas fluorescens, Serratia liquefaciens, Stenotrophomonas maltophilia). Despite hampered in growth in mixed cultures, L. monocytogenes established in biofilms with counts at day nine between 7.3 and 9.0 log per coupon with the lowest counts in the Listeria + G- mix that was dominated by Pseudomonas. Specific L. innocua inhibited growth of L. monocytogenes strains differently; inhibition that was further enhanced by the background Gram-negative microbiota. In these multispecies and multibacteria cultures, the growth competitive effects lead to the dominance of a strong competitor L. monocytogenes strain that was only slightly inhibited by L. innocua and showed strong competitive abilities in mixed cultures with resident Gram-negative bacteria. The results indicates complex patterns of bacterial interactions and L. monocytogenes inhibition in the multibacteria cultures that only partially depend on cell contact and likely involve various antagonistic and bacterial tolerance mechanisms. The study indicates large variations among L. monocytogenes in their competitiveness under multibacterial culture conditions that should be considered in further studies towards understanding of L. monocytogenes persistence in food processing facilities.


Reduced mature microRNA levels in association with dicer loss in human temporal lobe epilepsy with hippocampal sclerosis.
Hippocampal sclerosis (HS) is a common pathological finding in patients with temporal lobe epilepsy (TLE) and is associated with altered expression of genes controlling neuronal excitability, glial function, neuroinflammation and cell death. MicroRNAs (miRNAs), a class of small non-coding RNAs, function as post-transcriptional regulators of gene expression and are critical for normal brain development and function. Production of mature miRNAs requires Dicer, an RNAase III, loss of which has been shown to cause neuronal and glial dysfunction, seizures, and neurodegeneration. Here we investigated miRNA biogenesis in hippocampal and neocortical resection specimens from pharmacoresistant TLE patients and autopsy controls. Western blot analysis revealed protein levels of Dicer were significantly lower in certain TLE patients with HS. Dicer levels were also reduced in the hippocampus of mice subject to experimentally-induced epilepsy. To determine if Dicer loss was associated with altered miRNA processing, we profiled levels of 380 mature miRNAs in control and TLE-HS samples. Expression of nearly 200 miRNAs was detected in control human hippocampus. In TLE-HS samples there was a large-scale reduction of miRNA expression, with 51% expressed at lower levels and a further 24% not detectable. Primary transcript (pri-miRNAs) expression levels for several tested miRNAs were not different between control and TLE-HS samples. These findings suggest loss of Dicer and failure of mature miRNA expression may be a feature of the pathophysiology of HS in patients with TLE.


Effects of silicon, citrate and the fasting state on the intestinal absorption of aluminium in rats.
1. The effect of silicon (Si) contained in drinking water and solid food on the intestinal absorption of aluminium (Al) remains a matter of debate. The present study was designed to readdress this issue in the experimental animal, and to examine concomitantly the effects of citrate and the fasting state, respectively. 2. Three groups of young, non-fasted rats (n = 8 per group) were gavaged by solutions containing 3.8 ng of 26Al, 63 ng of 27Al, and either distilled water (< 0.1 mg/l Si) or commercial mineral water with a medium (6 mg/l) or high (14 mg/l) Si concentration. 3. Two other groups of eight non-fasted rats each received the same distilled water or high-Si gavage solution, respectively, together with a high citrate concentration (62 g/l). In each case the animals had free access to drinking water for 5 days before and 2 days after the gavage, containing the same Si concentration as in the gavage solution. A sixth group of eight rats was gavaged by low-Si, Al and distilled water in the fasted state. 4. The animals were killed 48 h after gavage, and blood, tissue and urine samples were collected for 26Al measurements by accelerator mass spectrometry. 5. We found that the fraction of absorbed 26Al retained in the skeleton (0.025-0.030%) was of the same order of magnitude as the fraction excreted in the 48 h urine (0.035-0.037%). High Si concentrations in the drinking water failed to depress the 26Al fraction absorbed, as estimated on the basis of skeletal accumulation and urinary excretion. 6. The administration of citrate-containing fluid enhanced 26Al absorption 5- to 10-fold (P < 0.005), but again the Si content of drinking water did not interfere. Finally, the intestinal absorption of 26Al was approximately 15 times higher in the fasted than in the non-fasted state. 7. In conclusion, the provision of large amounts of Si in the drinking water failed to modify physiological intestinal Al absorption under basal conditions or after its stimulation by citrate. However, a prolonged fast greatly enhanced Al absorption, compared with the non-fasted state.


An ancient dental gene set governs development and continuous regeneration of teeth in sharks.
The evolution of oral teeth is considered a major contributor to the overall success of jawed vertebrates. This is especially apparent in cartilaginous fishes including sharks and rays, which develop elaborate arrays of highly specialized teeth, organized in rows and retain the capacity for life-long regeneration. Perpetual regeneration of oral teeth has been either lost or highly reduced in many other lineages including important developmental model species, so cartilaginous fishes are uniquely suited for deep comparative analyses of tooth development and regeneration. Additionally, sharks and rays can offer crucial insights into the characters of the dentition in the ancestor of all jawed vertebrates. Despite this, tooth development and regeneration in chondrichthyans is poorly understood and remains virtually uncharacterized from a developmental genetic standpoint. Using the emerging chondrichthyan model, the catshark (Scyliorhinus spp.), we characterized the expression of genes homologous to those known to be expressed during stages of early dental competence, tooth initiation, morphogenesis, and regeneration in bony vertebrates. We have found that expression patterns of several genes from Hh, Wnt/β-catenin, Bmp and Fgf signalling pathways indicate deep conservation over ~450 million years of tooth development and regeneration. We describe how these genes participate in the initial emergence of the shark dentition and how they are redeployed during regeneration of successive tooth generations. We suggest that at the dawn of the vertebrate lineage, teeth (i) were most likely continuously regenerative structures, and (ii) utilised a core set of genes from members of key developmental signalling pathways that were instrumental in creating a dental legacy redeployed throughout vertebrate evolution. These data lay the foundation for further experimental investigations utilizing the unique regenerative capacity of chondrichthyan models to answer evolutionary, developmental, and regenerative biological questions that are impossible to explore in classical models.


Fracture mechanics of delamination defects in multilayer dielectric coatings.
During the fabrication of multilayer-dielectric (MLD) thin-film-coated optics, such as the diffraction gratings used in OMEGA EP's pulse compressors, acid piranha cleaning can lead to the formation of chemically induced delamination defects. We investigate the causes of these defects and describe a mechanism for the deformation and failure of the MLD coating in response to hydrogen peroxide in the cleaning solution. A fracture mechanics model is developed and used to calculate the crack path that maximizes the energy-release rate, which is found to be consistent with the characteristic fracture pattern observed in MLD coating delamination defects.


Chemogenomic discovery of allosteric antagonists at the GPRC6A receptor.
GPRC6A is a Family C G protein-coupled receptor recently discovered and deorphanized by our group. This study integrates chemogenomic ligand inference, homology modeling, compound synthesis, and pharmacological mechanism-of-action studies to disclose two noticeable results of methodological and pharmacological character: (1) chemogenomic lead identification through the first, to our knowledge, ligand inference between two different GPCR families, Families A and C; and (2) the discovery of the most selective GPRC6A allosteric antagonists discovered to date. The unprecedented inference of pharmacological activity across GPCR families provides proof-of-concept for in silico approaches against Family C targets based on Family A templates, greatly expanding the prospects of successful drug design and discovery. The antagonists were tested against a panel of seven Family A and C G protein-coupled receptors containing the chemogenomic binding sequence motif where some of the identified GPRC6A antagonists showed some activity. However, three compounds with at least ∼3-fold selectivity for GPRC6A were discovered, which present a significant step forward compared with the previously published GPRC6A antagonists, calindol and NPS 2143, which both display ∼30-fold selectivity for the calcium-sensing receptor compared to GPRC6A. The antagonists constitute novel research tools toward investigating the signaling mechanism of the GPRC6A receptor at the cellular level and serve as initial ligands for further optimization of potency and selectivity enabling future ex vivo/in vivo pharmacological studies.


Cell-cell contact viral transfer contributes to HIV infection and persistence in astrocytes.
Astrocytes are the most abundant cells in the central nervous system and play important roles in human immunodeficiency virus (HIV)/neuro-acquired immunodeficiency syndrome. Detection of HIV proviral DNA, RNA, and early gene products but not late structural gene products in astrocytes in vivo and in vitro indicates that astrocytes are susceptible to HIV infection albeit in a restricted manner. We as well as others have shown that cell-free HIV is capable of entering CD4- astrocytes through human mannose receptor-mediated endocytosis. In this study, we took advantage of several newly developed fluorescence protein-based HIV reporter viruses and further characterized HIV interaction with astrocytes. First, we found that HIV was successfully transferred to astrocytes from HIV-infected CD4+ T cells in a cell-cell contact- and gp120-dependent manner. In addition, we demonstrated that, compared to endocytosis-mediated cell-free HIV entry and subsequent degradation of endocytosed virions, the cell-cell contact between astrocytes and HIV-infected CD4+ T cells led to robust HIV infection of astrocytes but retained the restricted nature of viral gene expression. Furthermore, we showed that HIV latency was established in astrocytes. Lastly, we demonstrated that infectious progeny HIV was readily recovered from HIV latent astrocytes in a cell-cell contact-mediated manner. Taken together, our studies point to the importance of the cell-cell contact-mediated HIV interaction with astrocytes and provide direct evidence to support the notion that astrocytes are HIV latent reservoirs in the central nervous system.


Developmental effects in fish embryos exposed to oil dispersions - The impact of crude oil micro-droplets.
During accidental crude oil spills and permitted discharges of produced water into the marine environment, a large fraction of naturally occurring oil components will be contained in micron-sized oil droplets. Toxicity is assumed to be associated with the dissolved fraction of oil components, however the potential contribution of oil droplets to toxicity is currently not well known. In the present work we wanted to evaluate the contribution of oil droplets to effects on normal development of Atlantic cod (Gadus morhua) through exposing embryos for 96 h to un-filtered (dispersions containing droplets) and filtered (water soluble fractions) dispersions in a flow-through system at dispersion concentrations ranging from 0.14 to 4.34 mg oil/L. After exposure, the embryos were kept in clean seawater until hatch when survival, development and morphology were assessed. The experiment was performed at two different stages of embryonic development to cover two potentially sensitive stages (gastrulation and organogenesis). Exposure of cod embryos to crude oil dispersions caused acute and delayed toxicity, including manifestation of morphological deformations in hatched larvae. Oil droplets appear to contribute to some of the observed effects including mortality, larvae condition (standard length, body surface, and yolk sac size), spinal deformations as well as alterations in craniofacial and jaw development. The timing of exposure may be essential for the development of effects as higher acute mortality was observed when embryos were exposed from the start of gastrulation (Experiment 1) than when exposed during organogenesis (Experiment 2). Even though low mortality was observed when exposed during organogenesis, concentration-dependent mortality was observed during recovery.


Antifungal prophylaxis in newly diagnosed AML patients-Adherence to guidelines and feasibility in a real life setting.
Antifungal posaconazole prophylaxis for AML patients receiving induction chemotherapy has been routine at our centre since 2009. This retrospective study examined the feasibility and practicability of our prophylaxis guidelines in clinical practice. Data sets of 90 patients undergoing induction-chemotherapy for AML between 2011 and 2014 were evaluated regarding adherence to local guidelines for the administration of antifungal prophylaxis with posaconazole. 75.5% of the 90 patients received posaconazole prophylaxis. All but eight patients received the recommended dosage. A total of 77.95% on prophylaxis had serum galactomannan measured twice weekly. Contradicting our guidelines, 89.70% of patients received concomitant therapy with PPI. Overall, 16.17% of patients had prophylaxis discontinued and started empirical antifungal treatment in the absence of diagnostic criteria for IFI. The breakthrough IFI rate was 36.76% (proven, probable and possible) with 7.35% of infections being classified as proven or probable. Although limited by a small sample size, our study demonstrates the feasibility of local guidelines in a real life setting and outlines areas for improvement in both guidelines and clinical practice. We also highlight the importance of ensuring awareness of guidelines and raise questions about a uniform approach to antifungal prophylaxis in AML patients.


Space-time clustering of recently-diagnosed tuberculosis and impact of ART scale-up: Evidence from an HIV hyper-endemic rural South African population.
In HIV hyperendemic sub-Saharan African communities, particularly in southern Africa, the likelihood of achieving the Sustainable Development Goal of ending the tuberculosis (TB) epidemic by 2030 is low, due to lack of cost-effective and practical interventions in population settings. We used one of Africa's largest population-based prospective cohorts from rural KwaZulu-Natal Province, South Africa, to measure the spatial variations in the prevalence of recently-diagnosed TB disease, and to quantify the impact of community coverage of antiretroviral therapy (ART) on recently-diagnosed TB disease. We collected data on TB disease episodes from a population-based sample of 41,812 adult individuals between 2009 and 2015. Spatial clusters ('hotspots') of recently-diagnosed TB were identified using a space-time scan statistic. Multilevel logistic regression models were fitted to investigate the relationship between community ART coverage and recently-diagnosed TB. Spatial clusters of recently-diagnosed TB were identified in a region characterized by a high prevalence of HIV and population movement. Every percentage increase in ART coverage was associated with a 2% decrease in the odds of recently-diagnosed TB (aOR = 0.98, 95% CI:0.97-0.99). We identified for the first time the clear occurrence of recently-diagnosed TB hotspots, and quantified potential benefit of increased community ART coverage in lowering tuberculosis, highlighting the need to prioritize the expansion of such effective population interventions targeting high-risk areas.


Sex determination in Drosophila melanogaster and Musca domestica converges at the level of the terminal regulator doublesex.
Sex-determining cascades are supposed to have evolved in a retrograde manner from bottom to top. Wilkins' 1995 hypothesis finds support from our comparative studies in Drosophila melanogaster and Musca domestica, two dipteran species that separated some 120 million years ago. The sex-determining cascades in these flies differ at the level of the primary sex-determining signal and their targets, Sxl in Drosophila and F in Musca. Here we present evidence that they converge at the level of the terminal regulator, doublesex ( dsx), which conveys the selected sexual fate to the differentiation genes. The dsx homologue in Musca, Md-dsx, encodes male-specific (MdDSX(M)) and female-specific (MdDSX(F)) protein variants which correspond in structure to those in Drosophila. Sex-specific regulation of Md-dsx is controlled by the switch gene F via a splicing mechanism that is similar but in some relevant aspects different from that in Drosophila. MdDSX(F) expression can activate the vitellogenin genes in Drosophila and Musca males, and MdDSX(M) expression in Drosophila females can cause male-like pigmentation of posterior tergites, suggesting that these Musca dsx variants are conserved not only in structure but also in function. Furthermore, downregulation of Md-dsx activity in Musca by injecting dsRNA into embryos leads to intersexual differentiation of the gonads. These results strongly support a role of Md-dsx as the final regulatory gene in the sex-determining hierarchy of the housefly.


Compensatory strategies below the behavioural surface in autism: a qualitative study.
Little is known about the compensatory profile in autism; that is, people with autism spectrum disorder who show few symptoms in their behavioural presentation, despite continuing to report autism-related cognitive difficulties or differences. Even less is known about the specific compensatory strategies that these individuals use to disguise autism at the behavioural surface, both in the clinic and everyday life. It is also currently unclear whether individuals without a formal autism diagnosis, but experiencing autistic-like difficulties, use similar compensatory strategies, potentially enabling them to sit below the diagnostic threshold. This study aimed to investigate social compensatory strategies, and their effect on diagnosis and clinical outcome, in adults with and without autism. In this study, individuals aged 18 years or older who responded to a study advert that was distributed worldwide via social media and the UK National Autistic Society formed a convenience sample. Participants self-reported their use and experiences of compensatory strategies using an online platform. Novel analyses, including a qualitative thematic approach, were used to interpret their responses and gain insight into compensatory strategies in autism. Between Oct 19, 2017, and Jan 2, 2018, 136 adults (58 had a clinical diagnosis of autism, 19 self-identified but were not formally diagnosed as autistic, and 59 were not diagnosed or self-identified, but nevertheless reported social difficulties) completed the online study questions. The findings suggested that there are multiple compensatory strategies with distinct characteristics, individual and environmental factors that modulate compensatory strategy use and success, positive (social relationships, independence, employment) and negative (poor mental health, late diagnosis) outcomes associated with compensatory strategy use, and that individuals without a diagnosis use compensatory strategies that are qualitatively similar to individuals with a diagnosis. Increased awareness and measurement of compensatory strategy use in autism should guide future diagnostic guidelines, towards improved diagnostic accuracy and support for people with autism spectrum disorder whose cognitive difficulties are not immediately evident in observable behaviour. UK Medical Research Council and UK National Institute for Health Research.


The D816V mutation of c-Kit circumvents a requirement for Src family kinases in c-Kit signal transduction.
The receptor tyrosine kinase c-Kit plays a critical role in hematopoiesis, and gain-of-function mutations of the receptor are frequently seen in several malignancies, including acute myeloid leukemia, gastrointestinal stromal tumors, and testicular carcinoma. The most common mutation of c-Kit in these disorders is a substitution of the aspartic acid residue in position 816 to a valine (D816V), leading to constitutive activation of the receptor. In this study, we aimed to investigate the role of Src family kinases in c-Kit/D816V signaling. Src family kinases are necessary for the phosphorylation of wild-type c-Kit as well as of activation of downstream signaling pathways including receptor ubiquitination and the Ras/Mek/Erk pathway. Our data demonstrate that, unlike wild-type c-Kit, the phosphorylation of c-Kit/D816V is not dependent on Src family kinases. In addition, we found that neither receptor ubiquitination nor Erk activation by c-Kit/D816V required activation of Src family kinases. In vitro kinase assay using synthetic peptides revealed that c-Kit/D816V had an altered substrate specificity resembling Src and Abl tyrosine kinases. We further present evidence that, in contrast to wild-type c-Kit, Src family kinases are dispensable for c-Kit/D816V cell survival, proliferation, and colony formation. Taken together, we demonstrate that the signal transduction pathways mediated by c-Kit/D816V are markedly different from those activated by wild-type c-Kit and that altered substrate specificity of c-Kit circumvents a need for Src family kinases in signaling of growth and survival, thereby contributing to the transforming potential of c-Kit/D816V.


Implementing service improvement projects within pre-registration nursing education: a multi-method case study evaluation.
Preparing healthcare students for quality and service improvement is important internationally. A United Kingdom (UK) initiative aims to embed service improvement in pre-registration education. A UK university implemented service improvement teaching for all nursing students. In addition, the degree pathway students conducted service improvement projects as the basis for their dissertations. The study aimed to evaluate the implementation of service improvement projects within a pre-registration nursing curriculum. A multi-method case study was conducted, using student questionnaires, focus groups with students and academic staff, and observation of action learning sets. Questionnaire data were analysed using SPSS v19. Qualitative data were analysed using Ritchie and Spencer's (1994) Framework Approach. Students were very positive about service improvement. The degree students, who conducted service improvement projects in practice, felt more knowledgeable than advanced diploma students. Selecting the project focus was a key issue and students encountered some challenges in practice. Support for student service improvement projects came from action learning sets, placement staff, and academic staff. Service improvement projects had a positive effect on students' learning. An effective partnership between the university and partner healthcare organisations, and support for students in practice, is essential.


Mortality experience of employees exposed to 2-mercaptobenzothiazole at a chemical plant in Nitro, West Virginia.
Mortality trends for 1059 production workers at a rubber chemicals plant in Nitro, West Virginia were examined to find whether they had increased mortality from cancer associated with exposure to 2-mercaptobenzothiazole (MBT). This chemical and its derivatives are vulcanising agents that have been manufactured at the plant since 1935. Analyses were conducted on MBT exposed employees by cumulative exposure and time since first exposure, and were also stratified by past assignment to p-aminobiphenyl (PAB) related departments; PAB is a potent bladder carcinogen that was used at the plant between 1935 and 1955. There was an excess of bladder cancer in MBT workers who had PAB related assignments (standardised mortality ratio (SMR) = 3200, 95% confidence interval (95% CI) 1286-6593). In employees without a job assignment with exposure to PAB, there were no associations between exposure to MBT and increased rates of most malignant neoplasms. The SMR for bladder cancer was increased based on three deaths (SMR = 455, 95% CI 94-1328), although these results were too few to evaluate trends by cumulative exposure category. The possibility of confounding by PAB for exposures for jobs that covered all areas of the plant for these three cases must be considered in the light of the potency of PAB as a bladder carcinogen. There were no deaths from bladder cancer among MBT workers hired after the end of manufacture and use of PAB, but the expected number of deaths was only 0.03.


[The effects of a Korean traditional dance movement program in elderly women].
This study was aimed to identify the effects of a 12 week Korean traditional dance movement program on balance, depression, medical cost, medical institution's utilization and fall among elderly women. Using a quasi-experimental design, the experimental group was composed of 130 subjects and the control group was composed of 123 subjects. The experimental group participated in a 12 week Korean traditional dance movement program 3 times a week from December 2002 to February 2003. Data was analyzed with descriptive statistics, the chi-square test, paired t-test and t-test. There was significant improvement in balance (right leg p=.000, left leg p=.004), depression (p=.000), and the medical institution's utilization (p=.001) and fall (p=.002) in the experimental group compared to the control group. A Korean traditional dance movement program improved balance, depression, and decreased fall and medical cost in elderly women. Therefore, we recommend this program be utilized as a health promoting program and falls preventing program for the elderly in the community.


The use of sural island musculo fasciocutaneous flap, in the management of chronic osteomyelitis of the tibia.
: The management of soft-tissue defects following surgery for chronic osteomyelitis of the tibia is challenging. It often requires complex reconstructive procedures, especially when the distal third of the tibia is involved. We present a relatively simple method of addressing these defects. : This study aims to report our experience with the use of the sural Island musculo fasciocutaneous flap in the management of osteomyelitis of the tibia. : Consecutive patients with osteomyelitis of the tibia requiring soft-tissue cover were managed using the sural Island musculo fasciocutaneous flap, and the outcome was analyzed. The study was prospective in nature. : There were 21 patients, 15 males and 6 females. Their ages ranged from 21 years to 62 years. Most (71%) were in the 3rd and 4th decades of life. Twelve patients had involvement of the distal third of the tibia, eight involved the middle third, while five patients had involvement of the proximal third. Most of the patients (17) developed osteomyelitis following open fractures of the tibia; thirteen of these were from road crashes and four from gunshot injuries. Four patients developed osteomyelitis through the hematogenous route. Flap sizes ranged from 8 × 7 cm to 16 × 11 cm. There was tip necrosis in 2 flaps and full survival in 19 flaps. The flaps with partial necrosis were managed by simple dressing. Fifteen patients (71%) had cessation of drainage within 4 weeks of surgery. Two patients presented with recurrence of infection within the period of follow-up. Mean follow-up period was 23 months. All donor sites were skin grafted primarily, with 95%-100% graft take. The sural Island musculo fasciocutaneous flap is a reliable source of richly vascularized soft tissue for the management of dead space and soft-tissue defects in chronic osteomyelitis of the tibia. It has the added advantage of providing sizeable tissue, with good reach to all segments of the tibia.


Growth substrate- and phase-specific expression of biphenyl, benzoate, and C1 metabolic pathways in Burkholderia xenovorans LB400.
Recent microarray experiments suggested that Burkholderia xenovorans LB400, a potent polychlorinated biphenyl (PCB)-degrading bacterium, utilizes up to three apparently redundant benzoate pathways and a C(1) metabolic pathway during biphenyl and benzoate metabolism. To better characterize the roles of these pathways, we performed quantitative proteome profiling of cells grown on succinate, benzoate, or biphenyl and harvested during either mid-logarithmic growth or the transition between the logarithmic and stationary growth phases. The Bph enzymes, catabolizing biphenyl, were approximately 16-fold more abundant in biphenyl- versus succinate-grown cells. Moreover, the upper and lower bph pathways were independently regulated. Expression of each benzoate pathway depended on growth substrate and phase. Proteins specifying catabolism via benzoate dihydroxylation and catechol ortho-cleavage (ben-cat pathway) were approximately an order of magnitude more abundant in benzoate- versus biphenyl-grown cells at the same growth phase. The chromosomal copy of the benzoyl-coenzyme A (CoA) (box(C)) pathway was also expressed during growth on biphenyl: Box(C) proteins were approximately twice as abundant as Ben and Cat proteins under these conditions. By contrast, proteins of the megaplasmid copy of the benzoyl-CoA (box(M)) pathway were only detected in transition-phase benzoate-grown cells. Other proteins detected at increased levels in benzoate- and biphenyl-grown cells included general stress response proteins potentially induced by reactive oxygen species formed during aerobic aromatic catabolism. Finally, C(1) metabolic enzymes were present in biphenyl-grown cells during transition phase. This study provides insights into the physiological roles and integration of apparently redundant catabolic pathways in large-genome bacteria and establishes a basis for investigating the PCB-degrading abilities of this strain.


Adherence monitoring of rehabilitation exercise with inertial sensors: A clinical validation study.
Rehabilitation has an established role in the management of a wide range of musculoskeletal conditions. Much of this treatment relies on self-directed exercises at home, where adherence of execution is unknown. Demonstrating treatment fidelity is necessary to draw conclusions about the efficacy of rehabilitation interventions in both clinical and research settings. There is a lack of tools and methods to achieve this. This study aims to evaluate the feasibility of using a single inertial sensor to recognise and classify shoulder rehabilitation activity using supervised machine learning techniques. Twenty patients with shoulder pain were monitored performing five rehabilitation exercises routinely prescribed for their condition. Accelerometer, gyroscope and magnetometer data were collected via a device mounted onto an arm sleeve. Non-specific motion data was included in the analysis. Time and frequency domain features were calculated from labelled data segments and ranked in terms of their predictive importance using the ReliefF algorithm. Selected features were used to train four supervised learning algorithms: decision tree, k-nearest neighbour, support vector machine and random forests. Performance of algorithms in accurately classifying exercise activity was evaluated with ten-fold cross-validation and leave-one-subject-out-validation methods. Optimal predictive accuracies for ten-fold cross-validation (97.2%) and leave-one-subject-out-validation (80.5%) were achieved by support vector machine and random forests algorithms, respectively. Time domain features derived from accelerometer, magnetometer and orientation data streams were shown to have the highest predictive value for classifying rehabilitation activity. Classification models performed well in differentiating patient exercise activity from non-specific movement and identifying specific exercise type using inertial sensor data. A clinically useful account of home rehabilitation activity will help guide treatment strategies and facilitate methods to improve patient engagement. Future work should focus on evaluating the performance of such systems in natural and unsupervised settings.


Impact of amoxicillin therapy on resistance selection in patients with community-acquired lower respiratory tract infections: a randomized, placebo-controlled study.
To determine the effect of amoxicillin treatment on resistance selection in patients with community-acquired lower respiratory tract infections in a randomized, placebo-controlled trial. Patients were prescribed amoxicillin 1 g, three times daily (n = 52) or placebo (n = 50) for 7 days. Oropharyngeal swabs obtained before, within 48 h post-treatment and at 28-35 days were assessed for proportions of amoxicillin-resistant (ARS; amoxicillin MIC ≥2 mg/L) and -non-susceptible (ANS; MIC ≥0.5 mg/L) streptococci. Alterations in amoxicillin MICs and in penicillin-binding-proteins were also investigated. ITT and PP analyses were conducted. ARS and ANS proportions increased 11- and 2.5-fold, respectively, within 48 h post-amoxicillin treatment compared with placebo [ARS mean increase (MI) 9.46, 95% CI 5.57-13.35; ANS MI 39.87, 95% CI 30.96-48.78; P < 0.0001 for both]. However, these differences were no longer significant at days 28-35 (ARS MI -3.06, 95% CI -7.34 to 1.21; ANS MI 4.91, 95% CI -4.79 to 14.62; P > 0.1588). ARS/ANS were grouped by pbp mutations. Group 1 strains exhibited significantly lower amoxicillin resistance (mean MIC 2.8 mg/L, 95% CI 2.6-3.1) than group 2 (mean MIC 9.3 mg/L, 95% CI 8.1-10.5; P < 0.0001). Group 2 strains predominated immediately post-treatment (61.07%) and although decreased by days 28-35 (30.71%), proportions remained higher than baseline (18.70%; P = 0.0004). By utilizing oropharyngeal streptococci as model organisms this study provides the first prospective, experimental evidence that resistance selection in patients receiving amoxicillin is modest and short-lived, probably due to 'fitness costs' engendered by high-level resistance-conferring mutations. This evidence further supports European guidelines that recommend amoxicillin when an antibiotic is indicated for community-acquired lower respiratory tract infections.


Instantaneous temperature imaging of diffusion flames using two-line atomic fluorescence.
This work investigates the first demonstration of nonlinear regime two-line atomic fluorescence (NTLAF) thermometry in laminar non-premixed flames. The results show the expediency of the technique in the study of the reaction zone and reveals interesting findings about the indium atomization process. Indium fluorescence is observed to be strongest at the flame-front, where the temperature exceeds 1000 K. The uncertainty in the deduced temperature measurement is approximately 6%. The temperature profile across the reaction zone shows good agreement with laminar flame calculations. The advantages and inherent limitations of the technique are discussed.


Modelling normal and impaired letter recognition: implications for understanding pure alexic reading.
Letter recognition is the foundation of the human reading system. Despite this, it tends to receive little attention in computational modelling of single word reading. Here we present a model that can be trained to recognise letters in various spatial transformations. When presented with degraded stimuli the model makes letter confusion errors that correlate with human confusability data. Analyses of the internal representations of the model suggest that a small set of learned visual feature detectors support the recognition of both upper case and lower case letters in various fonts and transformations. We postulated that a damaged version of the model might be expected to act in a similar manner to patients suffering from pure alexia. Summed error score generated from the model was found to be a very good predictor of the reading times of pure alexic patients, outperforming simple word length, and accounting for 47% of the variance. These findings are consistent with a hypothesis suggesting that impaired visual processing is a key to understanding the strong word-length effects found in pure alexic patients.


N-ethyl-N-Nitrosourea (ENU) induced mutations within the klotho gene lead to ectopic calcification and reduced lifespan in mouse models.
Ectopic calcification (EC), which is the pathological deposition of calcium and phosphate in extra-skeletal tissues, may be associated with hypercalcaemic and hyperphosphataemic disorders, or it may occur in the absence of metabolic abnormalities. In addition, EC may be inherited as part of several monogenic disorders and studies of these have provided valuable insights into the metabolic pathways regulating mineral metabolism. For example, studies of tumoural calcinosis, a disorder characterised by hyperphosphataemia and progressive EC, have revealed mutations of fibroblast growth factor 23 (FGF23), polypeptide N-acetyl galactosaminyltransferase 3 (GALNT3) and klotho (KL), which are all part of a phosphate-regulating pathway. However, such studies in humans are limited by the lack of available large families with EC, and to facilitate such studies we assessed the progeny of mice treated with the chemical mutagen N-ethyl-N-nitrosourea (ENU) for EC. This identified two mutants with autosomal recessive forms of EC, and reduced lifespan, designated Ecalc1 and Ecalc2. Genetic mapping localized the Ecalc1 and Ecalc2 loci to a 11.0 Mb region on chromosome 5 that contained the klotho gene (Kl), and DNA sequence analysis identified nonsense (Gln203Stop) and missense (Ile604Asn) Kl mutations in Ecalc1 and Ecalc2 mice, respectively. The Gln203Stop mutation, located in KL1 domain, was severely hypomorphic and led to a 17-fold reduction of renal Kl expression. The Ile604Asn mutation, located in KL2 domain, was predicted to impair klotho protein stability and in vitro expression studies in COS-7 cells revealed endoplasmic reticulum retention of the Ile604Asn mutant. Further phenotype studies undertaken in Ecalc1 (kl203X/203X) mice demonstrated elevations in plasma concentrations of phosphate, FGF23 and 1,25-dihydroxyvitamin D. Thus, two allelic variants of Kl that develop EC and represent mouse models for tumoural calcinosis have been established.


Neuromuscular blockade improves first-attempt success for intubation in the intensive care unit. A propensity matched analysis.
The use of neuromuscular blocking agents (NMBAs) has been shown to be valuable in improving successful tracheal intubation in the operating room and emergency department. However, data on NMBA use in critically ill intensive care unit (ICU) patients are lacking. Furthermore, there are no data on NMBA use with video laryngoscopy. To evaluate the effect of NMBA use on first-attempt success (FAS) with tracheal intubation in the ICU. Single-center observational study of 709 consecutive patients intubated in the medical ICU of a university medical center from January 1, 2012 to June 30, 2014. Data were collected prospectively through a continuous quality improvement program on all patients intubated in the ICU over the study period. Data relating to patient demographics, intubation, and complications were analyzed. We used propensity score (propensity to use an NMBA) matching to generate 5,000 data sets of cases (failed first intubation attempts) matched to controls (successful first attempts) and conditional logistic regression to analyze the results. There were no significant differences in patient demographics, except median total difficult airway characteristics were higher in the non-NMBA group (2 vs. 1, P < 0.001). There were significant differences in the sedative used between groups and the operator level of training. More patients who were given NMBAs received etomidate (83 vs. 35%) and more patients in the non-NMBA group received ketamine (39 vs. 9%) (P < 0.001). The FAS for NMBA use was 80.9% (401/496) compared with 69.6% (117/168) for non-NMBA use (P = 0.003). The summary odds ratio for FAS when an NMBA was used from the propensity matched analyses was 2.37 (95% confidence interval, 1.36-4.88). In the subgroup of patients intubated with a video laryngoscope, propensity-adjusted odds of FAS with the use of an NMBA was 2.50 (1.43-4.37; P < 0.001). There were no differences in procedurally related complications between groups. After controlling for potential confounders, this propensity-adjusted analysis demonstrates improved odds of FAS at intubation in the ICU with the use of an NMBA. This improvement in FAS is seen even with the use of a video laryngoscope.


BSD2 is a Rubisco-specific assembly chaperone, forms intermediary hetero-oligomeric complexes, and is nonlimiting to growth in tobacco.
The folding and assembly of Rubisco large and small subunits into L8 S8 holoenzyme in chloroplasts involves many auxiliary factors, including the chaperone BSD2. Here we identify apparent intermediary Rubisco-BSD2 assembly complexes in the model C3 plant tobacco. We show BSD2 and Rubisco content decrease in tandem with leaf age with approximately half of the BSD2 in young leaves (~70 nmol BSD2 protomer.m2 ) stably integrated in putative intermediary Rubisco complexes that account for <0.2% of the L8 S8 pool. RNAi-silencing BSD2 production in transplastomic tobacco producing bacterial L2 Rubisco had no effect on leaf photosynthesis, cell ultrastructure, or plant growth. Genetic crossing the same RNAi-bsd2 alleles into wild-type tobacco however impaired L8 S8 Rubisco production and plant growth, indicating the only critical function of BSD2 is in Rubisco biogenesis. Agrobacterium mediated transient expression of tobacco, Arabidopsis, or maize BSD2 reinstated Rubisco biogenesis in BSD2-silenced tobacco. Overexpressing BSD2 in tobacco chloroplasts however did not alter Rubisco content, activation status, leaf photosynthesis rate, or plant growth in the field or in the glasshouse at 20°C or 35°C. Our findings indicate BSD2 functions exclusively in Rubisco biogenesis, can efficiently facilitate heterologous plant Rubisco assembly, and is produced in amounts nonlimiting to tobacco growth.


Bacterial modulins: a novel class of virulence factors which cause host tissue pathology by inducing cytokine synthesis.
Cytokines are a diverse group of proteins and glycoproteins which have potent and wide-ranging effects on eukaryotic cell function and are now recognized as important mediators of tissue pathology in infectious diseases. It is increasingly recognized that for many bacterial species, cytokine induction is a major virulence mechanism. Until recent years, the only bacterial component known to stimulate cytokine synthesis was lipopolysaccharide (LPS). It is only within the past decade that it has been clearly shown that many components associated with the bacterial cell wall, including proteins, glycoproteins, lipoproteins, carbohydrates, and lipids, have the capacity to stimulate mammalian cells to produce a diverse array of cytokines. It has been established that many of these cytokine-inducing molecules act by mechanisms distinct from that of LPS, and thus their activities are not due to LPS contamination. Bacteria produce a wide range of virulence factors which cause host tissue pathology, and these diverse factors have been grouped into four families: adhesins, aggressins, impedins, and invasins. We suggest that the array of bacterial cytokine-inducing molecules represents a new class of bacterial virulence factor, and, by analogy with the known virulence families, we suggest the term "modulin" to describe these molecules, because the action of cytokines is to modulate eukaryotic cell behavior. This review summarizes our current understanding of cytokine biology in relation to tissue homeostasis and disease and concisely reviews the current literature on the cytokine-inducing molecules produced by gram-negative and gram-positive bacteria, with an emphasis on the cellular mechanisms responsible for cytokine induction. We propose that modulins, by controlling the host immune and inflammatory responses, maintain the large commensal flora that all multicellular organisms support.


Electrophysiological characteristics of complex fractionated electrograms and high frequency activity in atrial fibrillation.
It is unclear whether atrial substrate with complex fractionated electrograms (CFAEs) is related to arrhythmogenesis. This study aimed to investigate the electrophysiology in CFAE and high dominant frequency (DF) areas. Atrial fibrillation (AF) was induced by rapid atrial pacing in heart failure (HF) rabbits (4 weeks after coronary artery ligation). Real-time substrate mapping, multielectrode array, and monophasic action potential recordings were used to study areas of CFAE and DF. Conventional microelectrode and western blot were used to record the action potentials (APs) and protein expression in isolated tissue preparations. CFAE site with high DF had the most depolarized resting membrane potential, highest incidence of early and delayed afterdepolarizations, and steepest maxima slope of 90% of AP duration (APD90) restitution curve (RC) compared to CFAE site with low DF or non-CFAE sites. CFAE site with high DF exhibited the slowest conduction velocity and shortest wavelength than the other areas. Upregulation of the Na(+)-Ca(2+) exchanger (NCX), apamin-sensitive small-conductance Ca(2+)-activated K(+) channel type 2 (SK2) and sarcoplasmic reticulum Ca(2+)-ATPase, and downregulation of the Kir2.1 were found at CFAE site with high DF compared to that observed in the 3 other areas. Inhibition of the NCX and SK channels prolonged the APD90, flattened the maximum slope of RC, and suppressed AF. CFAE site with high DF had an arrhythmogenic property differing significantly from the other areas of LA in an HF rabbit model, which may contribute to the genesis of AF.


Mechanism underlying long-term regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase during L6 myoblast differentiation.
3-Hydroxy 3-methylglutaryl Coenzyme A reductase (HMG-CoAR) and its end-products are crucial for insulin-induced differentiation of fetal rat myoblasts (L6) both at early and terminal stages of development. Inhibition of HMG-CoAR activity and reduction of the enzyme levels impair the expression of L6 differentiation markers and prevent myoblast fusion into multinucleated syncytia. The mechanism underlying the modulation of this crucial enzyme so that muscular differentiation can occur is poorly understood. Thus, the aim of this work was to explore the long-term regulation of HMG-CoAR in an attempt to provide a new molecular basis for the control of muscle development. All experiments were performed in L6 rat myoblasts induced to differentiate utilizing insulin. The results indicate the following: (i) at early stages of L6 differentiation, the increase in HMG-CoAR protein levels is probably due to transcriptional induction and a decrease in the enzyme degradation rate; (ii) the subsequent reduction of HMG-CoAR protein levels is related both to an increased degradation rate and reduced gene transcription, as indicated by the rise of Insig-1 levels and the subsequent decrease in the amount of n-SREBP-1; (iii) in the terminal stages of myogenesis, reduced protein levels of HMG-CoAR could be ascribed to the decrease in gene transcription while its degradation rate is not affected. By highlighting the mechanisms involved in HMG-CoAR long-term regulation during myogenesis, this work provides useful information for searching for tools to improve the regenerative ability of muscle tissue and for the development of new pharmacological treatments of myopathies.


Polyreactivity and autoreactivity among HIV-1 antibodies.
It is generally acknowledged that human broadly neutralizing antibodies (bNAbs) capable of neutralizing multiple HIV-1 clades are often polyreactive or autoreactive. Whereas polyreactivity or autoreactivity has been proposed to be crucial for neutralization breadth, no systematic, quantitative study of self-reactivity among nonneutralizing HIV-1 Abs (nNAbs) has been performed to determine whether poly- or autoreactivity in bNAbs is a consequence of chronic antigen (Ag) exposure and/or inflammation or a fundamental property of neutralization. Here, we use protein microarrays to assess binding to >9,400 human proteins and find that as a class, bNAbs are significantly more poly- and autoreactive than nNAbs. The poly- and autoreactive property is therefore not due to the infection milieu but rather is associated with neutralization. Our observations are consistent with a role of heteroligation for HIV-1 neutralization and/or structural mimicry of host Ags by conserved HIV-1 neutralization sites. Although bNAbs are more mutated than nNAbs as a group, V(D)J mutation per se does not correlate with poly- and autoreactivity. Infrequent poly- or autoreactivity among nNAbs implies that their dominance in humoral responses is due to the absence of negative control by immune regulation. Interestingly, four of nine bNAbs specific for the HIV-1 CD4 binding site (CD4bs) (VRC01, VRC02, CH106, and CH103) bind human ubiquitin ligase E3A (UBE3A), and UBE3A protein competitively inhibits gp120 binding to the VRC01 bNAb. Among these four bNAbs, avidity for UBE3A was correlated with neutralization breadth. Identification of UBE3A as a self-antigen recognized by CD4bs bNAbs offers a mechanism for the rarity of this bNAb class. Eliciting bNAbs is key for HIV-1 vaccines; most Abs elicited by HIV-1 infection or immunization, however, are strain specific or nonneutralizing, and unsuited for protection. Here, we compare the specificities of bNAbs and nNAbs to demonstrate that bNAbs are significantly more poly- and autoreactive than nNAbs. The strong association of poly- and autoreactivity with bNAbs, but not nNAbs from infected patients, indicates that the infection milieu, chronic inflammation and Ag exposure, CD4 T-cell depletion, etc., alone does not cause poly- and autoreactivity. Instead, these properties are fundamentally linked to neutralization breadth, either by the requirement for heteroligation or the consequence of host mimicry by HIV-1. Indeed, we show that human UBE3A shares an epitope(s) with HIV-1 envelope recognized by four CD4bs bNAbs. The poly- and autoreactivity of bNAbs surely contribute to the rarity of membrane-proximal external region (MPER) and CD4bs bNAbs and identify a roadblock that must be overcome to induce protective vaccines.


Formaldehyde exposure and respiratory and related cancers. A case-referent study among Finnish woodworkers.
A case-referent study was undertaken to investigate the associations between formaldehyde exposure and respiratory and related cancers. Fifty-seven such cancers from a retrospective cohort of male woodworkers formed the case group. They were matched by year of birth with 171 referents. Exposure to formaldehyde was assessed with job-exposure matrices. The median of the time-weighted average concentration was about 1 ppm, and the mean duration of exposure was 10 years among the exposed. Odds ratios (OR) were calculated for formaldehyde exposure (1.44), peak exposure to formaldehyde (1.26), and exposure to formaldehyde-containing wood dust (1.22). None of the values exceeded unity with statistical significance. Allowance was also made for a 10-year period from the onset of exposure. Birth year, cigarette smoking and exposure to wood dust, chlorophenols, pesticides, and terpenes were controlled by stratification. The adjusted ORs did not change appreciably. The highest OR was 1.95 for formaldehyde exposure without allowance for minimum latency, adjusted for exposure to terpenes. No exposure-response relation was observed for the level, duration, or dose (ppm-years) of formaldehyde exposure. The result is nonpositive and may be explained by absence of effect, by too short a follow-up, or by insufficient power for detecting a mild excess risk.